Cell Membrane ; metabolism ; Gastrointestinal Stromal Tumors ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Proto-Oncogene Proteins c-kit ; analysis

OBJECTIVETo investigate the morphological features, immunohistochemical speciality of the gastrointestinal stromal tumors (GISTs), and its histogenesis as well.

METHODSThe morphologic characteristics of GISTs were studied in 65 cases using light microscopy and 17 cases by electron microscopy. The expression of c-kit (CD117), CD34, vimentin, SMA, actin, desmin, S-100 and MBP were detected in all the cases with EnVision trade mark staining.

RESULTSAmong 65 cases of GISTs, 46 were spindle cell type, 6 epithelioid cell type and 13 mixture type, equivalent to 85.5% (65 of 76) of all of the mesenchymal tumors of gastrointestinal tract admitted in the same period. The epithelioid cell type tumors composed of mainly the epithelioid cells, predominantly short spindle, oval or round in pattern, with an overall eosinophilic cytoplasm by hematoxylin-eosin stain. Focal cytoplasmic vacuolization was often seen. Sometimes signet-ring like cells and cells with a clear cytoplasm were seen in the epithelioid stromal tumor. The tumor cells arranged in interlacing fascicles forming whorls or sometimes cell clusters. Electronic microscopy showed the presence of interdigitating cell processes, in some areas, synapse-like structure and numerous desmosome-like junctions as well as a few gap junctions and small round neurosecretory granules. There were also abundant intermediate filaments and thin filaments of actin-type with longitudinal condensations (dense bodies). All of the 65 stromal tumors were strongly positive for vimentin (100%), 61 out of 65 tumors positive for CD117 (c-kit) (93.8%) and 51 out of 65 positive for CD34 (78.5%). Some cases also expressed SMA, actin, S-100 and MBP.

CONCLUSIONSGISTs were the most frequent mesenchymal tumor seen in the gastrointestinal tract. Under light microscope, the morphology of stromal tumors looks sometimes like a leiomyoma and Schwannoma. The application of immunohistochemical markers (particularly CD117 and CD34) and ultrastructural study are considered necessary for the differential diagnosis. GISTs may originate from the pluripotential precursor cells like the interstitial cells of Cajal.

Gastrointestinal Neoplasms ; Gastrointestinal Stromal Tumors ; Humans ; Immunohistochemistry ; Leiomyoma ; Microscopy, Electron ; Proto-Oncogene Proteins c-kit ; metabolism

Mechanism study was performed to explore how Shouhui Tongbian Capsules promotes energy metabolism of gastrointestinal stromal cells. In this study, gastrointestinal stromal cells line GIST-882 was used as the model to explore energy metabolism regulation effects of Shouhui Tongbian Capsules extract(10, 20, 50 and 100 μg·mL~(-1)) by measuring the cell proliferation, ATP level, mitochondrial membrane potential, and mitochondrial isocitrate dehydrogenase activity. Meanwhile, Western blot was used to detect the proteins expression of SCF/c-Kit and CDK2/cyclin A signaling pathways. Our results showed that Shouhui Tongbian Capsules promoted cell proliferation and increased ATP level of gastrointestinal stromal cells. In addition, Shouhui Tongbian Capsules obviously improved mitochondrial structural integrity, and increased mitochondrial membrane potential in GIST-882 cells. Mechanism study revealed that Shouhui Tongbian Capsules increased mitochondrial isocitrate dehydrogenase activity and up-regulated the proteins expression of SCF/c-Kit and CDK2/cyclin A signaling pathways. Collectively, our study indicated that Shouhui Tongbian Capsules promoted the energy metabolism for gastrointestinal stromal cells proliferation by activating mitochondrial isocitrate dehydrogenase to induce ATP production, as well as activating SCF/c-Kit and CDK2/cyclin A signaling pathways.
Capsules ; Cell Line, Tumor ; Energy Metabolism ; Gastrointestinal Stromal Tumors ; Humans ; Proto-Oncogene Proteins c-kit/metabolism* ; Stromal Cells/metabolism*

Adult ; Exons ; Female ; Gastrointestinal Stromal Tumors ; genetics ; metabolism ; pathology ; Gene Deletion ; Humans ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism

OBJECTIVE: To investigate the safety of the controllable ileostomy with pipe in view of histology. METHODS: Twenty-eight Beagle dogs undergoing controllable ileostomy with pipe were studied. The special fistula tube with balloon was placed into the hole locating at the cecal root opposing the mesenteric side, and fixed by double knot compression method. RESULTS: The fistula tube was removed 14 days after surgery, then the safety of the procedure was preliminarily evaluated by gastrointestinal radiography and anatomical observation. The small intestine tissue at the compression suture was used as the experimental segment, and the small intestine tissue at the proximal non-compression suture was used as the control segment. The histological staining and the immunohistochemical staining of S-100 protein, c-kit protein and α-smooth muscle actin(α-SMA) protein between two segment were compared, while quantitative comparison of myenteric plexus, intestinal Cajal cell(ICC) and smooth muscle cells in intestinal wall was carried out. After removal of fistula tube at 14 days postoperative, the dogs were normal in feeding and defecation. The digestive tract radiography showed that the intestine was patent without obvious stenosis and obstruction. The dogs were dissected 21 days after operation. The abdominal sinus ostium was well healed and the internal sinus was well formed. Under gross inspection, blood supply, morphology and motor function of experimental intestine segment were similar from the proximal and distal segments of control intestine. S-100 immunohistochemical staining showed that the morphology and distribution of S-100 protein positive cells and "blank area" cells in the experimental and control segments were consistent. Myenteric plexus counting showed that the experimental segment was 3.62±1.82/field and the control segment was 3.27±1.62/field, whose difference was not statistically significant(t=1.30, P=0.20). Immunohistochemical staining of c-kit showed that the distribution of c-kit positive cells in both segments was consistent. Counting of the number of ICCs in myenteric plexus revealed that experimental segment was 2.96±2.57/plexus, and control segment was 2.49±1.80/plexus without significant difference(t=1.81, P=0.07). Immunohistochemical staining of α-SMA showed that the morphology and distribution of smooth muscle cells in whole intestinal wall(muscle layer, longitudinal muscle, ring muscle) in experimental and control segments were consistent. The average absorbance(A) value of α-SMA staining in ring muscle layer was detected and quantified. The experimental segment was 0.15±0.03 and control segment was 0.14±0.04 without significant difference(t=1.16, P=0.25). CONCLUSION The technique of controllable ileostomy with pipe is safe in view of histology, which may replace the traditional protective ileostomy.
Animals ; Dogs ; Ileostomy ; methods ; standards ; Intestine, Small ; surgery ; Models, Animal ; Proto-Oncogene Proteins c-kit ; metabolism ; Treatment Outcome

The aim of this study was to investigate the effect of microRNA-193b (miR-193b) on C-KIT protein expression and biological behaviors in K562 cells. The FAM-labeled miR-193b mimic and negative control were respectively transfected into K562 cells using HiPerFect transfection reagent. The percentage of FAM-positive cells was monitored by flow cytometry. The levels of C-KIT and phosphorylated C-KIT protein were detected by Western blot. The cell growth was measured by CCK-8 reagent. The apoptosis of cells were analyzed by flow cytometry with Annexin V staining. The differentiation of cells were analyzed by flow cytometry with anti-CD11b or anti-CD15 staining. The results demonstrated that the percentage of FAM-positive cells was about 80% in miR-193b or negative control-transfected K562 cells. Compared with the negative control group, overexpression of miR-193b in K562 cells significantly inhibited the levels of C-KIT and phosphorylated C-KIT protein. Meanwhile, the cell growth was inhibited and the percentages of apoptotic cells, CD11b- or CD15-positive cells increased. It is concluded that miR-193b can reduce C-KIT expression and inhibit cell growth in K562 cells. The growth-inhibitory activity of miR-193b is associated with apoptosis and granulocytic differentiation. This study contributed to further investigate the role of miR-193b in leukemogenesis.
Cell Differentiation ; genetics ; Cell Proliferation ; Humans ; K562 Cells ; MicroRNAs ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Transfection

Animals ; Bufo bufo ; Caspase 3 ; genetics ; metabolism ; Female ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the clinicopathologic features of gastrointestinal stromal tumors (GISTs) and to identify reliable prognostic parameters.

METHODSFifty-nine GISTs were studied by immunostaining of CD117, CD34, SMA, desmin, S-100, bcl-2, and Ki-67. Histopathologic evaluations included tumor size, necrosis, histological growth patterns, mitotic activities and tumor lymphocytic infiltrate. The patients were clinically followed for 2 to 9 years. Univariate, multivariate and correlative statistical evaluations were used to analyze the data.

RESULTSAmong the 59 patients, 40 were alive and 15 died of their tumors at follow-up, the remaining 4 patients died of other causes. Pathological parameters that correlated with prognosis included tumor sizes of more than 5 cm, tumor tissue necrosis, mitotic cell count equal or higher than 5 per 50 high power field, Ki-67 labeling index (LI) equal or higher than 5% and intense bcl-2 immunostaining. Multivariate analysis showed that the mitotic count and Ki-67 LI were independent prognostic indicators. There was a correlation between mitotic count and Ki-67 LI.

CONCLUSIONSMitotic count and Ki-67 LI are the best predictors for a poor outcome of GIST after surgical treatment. Ki-67 immunostaining may substitute mitotic count as a useful prognostic parameter.

Adolescent ; Adult ; Aged ; Child ; Cyclin D1 ; metabolism ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics

OBJECTIVETo examine the differences in the expressions of c-kit, HIWI and vimentin in the testis and epididymis of rats aged different months, investigate the regular changes in the expression of characteristic molecules in the ripening and senescent process of the testis and epididymis, and provide experimental evidence for studies on the aging of the male reproductive system.

METHODSSprague-Dawley male rats were divided into a young group (6-month-old, n=10), an adult group (12-month-old, n=10) and an aged group (24-month-old, n=10). The immunohistochemical SP method was used to examine c-kit, HIWI and vimentin in the testis and epididymis of the rats.

RESULTSThe positive immunohistochemical reaction to c-kit was observed mainly in spermatogonia, weakly in other cells and also in the epithelia and lumens of the epididymis. The abundant expression of HIWI was detected in all stages of spermatogenic cells in the testis and epididymis of the 6-month-old and 12-month-old rats. Moreover, spermatogonias and primary spermatocytes displayed intense expression. Sertoli cells, Leydig cells, myoid cells and vascular endothelial cells (VEC) were also HIWI-positive cells. But the positive expression decreased with age in the testis and epididymis of the 24-month-old rats (P > 0.05). vimentin expression was weak in the testis and epididymis of the 6-month-olds but increased significantly in the 24-month-olds (P > 0.01).

CONCLUSIONThe expressions of c-kit and HIWI decreased in the testis and epididymis of the 24-month-old rats, while vimentin expression increased markedly with age. The results suggest that the aging of the testis and epididymis is closely related to the abnormal transduction of cell signals.

Age Factors ; Animals ; Epididymis ; metabolism ; Immunohistochemistry ; Male ; Proteins ; metabolism ; Proto-Oncogene Proteins c-kit ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism ; Vimentin ; biosynthesis